Testosterone derivatives and assay method

Abstract

An anti-testosterone serum is provided which is highly selective to the measurement of testosterone using radioimmunossay techniques in both male and female plasma without resorting to chromatography even though the plasma may have a high 5α-dihydrotestosterone content. In addition, the high progesterone concentration normally present in female plasma has no effect on the assay of this plasma. The preferred hapten for producing the serum is 17β, 19-dihydroxyandrost-4-en-3-one 19-carboxymethyl ether although related compounds as disclosed herein are also useful.

The invention having been described, what is claimed is: 
     
       1. A compound selected from the group consisting of: ##STR9## wherein A is oxygen or sulfur; R is an alkyl group of 1 to 10 carbon atoms; and   B is one of (i) --OH, --OM where M is an alkali metal or an alkaline earth metal, or (ii) --OY where Y is an alkyl group of 1 to 10 carbon atoms, or (iii) an amide having the formula ##STR10##  where P is an alkyl group of 1 to 10 carbon atoms or a substituted aryl group having the general formula ##STR11##  with n being from 1 to 10 or a substituted heterocyclic ring with one or more nitrogen atoms in the ring and having the general formula ##STR12##  with n being from 1 to 10 and wherein the compounds are in the beta form.   
     
     
       2. The compound of claim 1 wherein A is oxygen. 
     
     
       3. The compound of claim 1 wherein R has one carbon atom. 
     
     
       4. The compound of claim 1 wherein B is --OH. 
     
     
       5. 17β, 19-dihydroxyandrost-4-en-3-one 19-carboxymethyl ether. 
     
     
       6. An immunogen comprising a compound of claim 1 conjugated to an immunological carrier. 
     
     
       7. An immunogen comprising a compound of claim 2 conjugated to an immunological carrier. 
     
     
       8. An immunogen comprising a compound of claim 3 conjugated to an immunological carrier. 
     
     
       9. An immunogen comprising a compound of claim 4 conjugated to an immunological carrier. 
     
     
       10. An immunogen comprising a compound of claim 5 conjugated to an immunological carrier. 
     
     
       11. The immunogen of claim 5 wherein the carrier is bovine serum albumin (BSA). 
     
     
       12. The antibody produced by injecting an immunogen of claim 6 into a host animal. 
     
     
       13. The antibody produced by injecting an immunogen of claim 7 into a host animal. 
     
     
       14. The antibody produced by injecting an immunogen of claim 8 into a host animal. 
     
     
       15. The antibody produced by injecting an immunogen of claim 9 into a host animal. 
     
     
       16. The antibody produced by injecting an immunogen of claim 10 into a host animal. 
     
     
       17. The antibody produced by injecting an immunogen of claim 11 into a host animal. 
     
     
       18. A method of making a radioimmunoassay of testosterone in a sample employing radiolabeled testosterone and an antibody, characterized by employing an antibody produced by injecting, into a host animal, a conjugate of 17β, 19-dihydroxyandrost-4-en-3-one 19-carboxymethyl ether and bovine serum albumin (BSA). 
     
     
       19. The method of claim 18 wherein the ether is thioether. 
     
     
       20. The method of claim 18 wherein the sample is prepared without chromatographic separation. 
     
     
       21. The method of making a radioimmunoassay of testosterone in a sample obtained from a female during her menstrual cycle so that the sample contains a relatively high content of progesterone as compared to the testosterone content comprising the step of conducting said assay using an antibody produced by injecting into a host animal, a conjugate of 17β, 19-dihydroxyandrost-4-en-3-one 19-carboxymethyl ether and bovine serum alubmin (BSA). 
     
     
       22. The method of claim 21 wherein the sample is prepared without chromatographic separation.